Abstract
Methods A phage library of 7000 clones was constructed from a drug naive HIV-1 clade C infected Indian patient whose plasma exhibited high potential neutralizing potential against a panel of viruses and also displayed cross-reactive anti-V3 antibodies. PBMCs were isolated and EBV transformed. Cells (wells) producing anti-V3 antibodies were preselected with V3-CTB fusion protein and expanded. Total RNA was isolated and cDNA was constructed followed by VH and VL amplification. scFvs were constructed, cloned into phagemid vector and expressed in Escherichia coli. We assessed the expression of the scFvs by SDSPAGE and Western blotting. Specificity was examined by ELISA.
Highlights
Open AccessGeneration and characterization of human monoclonal single chain variable fragments (scFvs) against envelope third variable region (V3) of HIV-1 clade C
Production of human monoclonal antibodies with broad neutralizing activity is an essential part of HIV-1 prophylactic vaccine
A phage library of 7000 clones was constructed from a drug naive HIV-1 clade C infected Indian patient whose plasma exhibited high potential neutralizing potential against a panel of viruses and displayed cross-reactive anti-V3 antibodies
Summary
Generation and characterization of human monoclonal single chain variable fragments (scFvs) against envelope third variable region (V3) of HIV-1 clade C. Rajesh Kumar1†, Raiees Andrabi1†, Ashutosh Tiwari, Somi Sankaran Prakash, Naveet Wig, Durgashree Dutta, Anurag Sankhyan, Lubina Khan, Subrata Sinha, Kalpana Luthra1*. From First International Science Symposium on HIV and Infectious Diseases (HIV SCIENCE 2012) Chennai, India. From First International Science Symposium on HIV and Infectious Diseases (HIV SCIENCE 2012) Chennai, India. 20-22 January 2012
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